Delivery systems for pharmacological agents encapsulated with proteinoids

ABSTRACT

Methods are described for targeting the release of an active pharmacological agent in an animal by administering that agent encapsulated in proteinoid microspheres which are stable to the environment encountered from the point of introduction until they migrate to the targeted body organs, fluids or cells and are there unstable. Orally administered delivery systems for insulin, heparin and physostigmine utilize encapsulating microspheres which are predominantly of less than about 10 microns in diameter and pass readily through the gastrointestinal mucosa and which are made of an acidic proteinoid that is stable and unaffected by stomach enzymes and acid, but which releases the microencapsulated agent in pharmacologically active form in the near neutral blood stream. Basic proteinoid microspheres encapsulating a dopamine redox carrier system are administered in the weakly basic, where they are stable, and then enter the blood stream, where the encapsulated agent is similarly released.

. .This is a continuation-in-part of copending application Ser. No.897,361, which was filed Aug. 18, 1986 and now is abandoned..!.

.Iadd.This is a continuation of application Ser. No. 07/883,562, filedMay 15, 1992 now abandoned; which is a reissue of application Ser. No.07/098,027, filed Sep. 8, 1987, now U.S. Pat. No. 4,925,673; which is acontinuation-in-part of application Ser. No. 06/897,361, filed Aug. 18,1986, now abandoned..Iaddend.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to pharmacologically active agents which areencapsulated within protective proteinoid microspheres and theadministration of same to warm blooded animals. It relates particularlyto orally administered microspheres containing pharmacological agentswhich otherwise would be deactivated in the gastrointestinal tract.

2. Description of the Prior Art

The available modes of delivery of pharmceutical and therapeutic agentsoften are severely limited by chemical or physical barriers or both,which are imposed by the body. For example, oral delivery of many suchagents would be the general method of choice if not for the numerousbarriers faced by these agents along this route. Gastrointestinalconditions of inappropriate pH, the presence of powerful digestiveenzymes, the permeability properties of gastrointestinal membranes andtissues and other factors all play important roles in determining thefeasibility of oral delivery of active agents to their targets. Amongthe numerous pharmacological agents which are known to be adverselyaffected or rendered ineffective when administered orally are thebiologically active polypeptides and proteins, such as insulin. Theseagents are rapidly destroyed in the stomach by acid hydrolysis and inthe stomach and lower gastrointestinal tract by enzymes capable ofcleaving peptide bonds and, in addition, they pass poorly, if at all,through the gastrointestinal wall.

A great deal of effort has been concentrated on the modification orisolation of the deleterious conditions within the gastrointestinaltract so that a pharmacological agent, which otherwise would be labile,could be absorbed through the stomach or intestine wall intact and inpharmacologically active form. The search in this area has been directedprimarily in three directions; the co-administration of adjuvants, suchas the resorcinols and the non-ionic surfactants polyoxyethylene oleylether and n-hexadecyl polyethylene ether; the co-administration ofenzymatic inhibitors, such as pancreatic trypsin inhibitor,diisopropylfluorophosphate (DFP) and trasylol; and the use of liposomes,such as water-in-oil-in-water emulsions which provide a protective layerof lipid around the encorporated pharmacological agent and whichrepresent the most successful approach to date. For example, the use ofliposomes containing heparin is disclosed in U.S. Pat. No. 4,239,754 andseveral studies have been directed to the use of liposomes containinginsulin; e.g., Patel et al, FEBS Letters, 62, 60 (1976) and Hashimoto etal, Endocrinol. Japan, 26, 337 (1979). In spite of these demonstrationsof limited operability, the use of liposomes is still in the developmentstage and there are continuing problems, including poor stability andinadequate shelf life.

Accordingly, there remains a need for improved means for targeting therelease of active pharmacological agents in the body and particularlyfor more satisfactory means for oral administration of pharmacologicalagents which are labile to conditions in the gastrointestinal tract.

SUMMARY OF THE INVENTION

It is an object of this invention to provide improved means forreleasing a pharmacological agent in physiologically active form at atargeted body organ or fluid.

It is a further object of this invention to provide an improved deliverysystem for the enteric administration or pharmacological agents which,by themselves, pass slowly or not at all through the gastro-intestinalmucosa and/or are susceptible to chemical cleavage by acids and enzymesin the gastrointestinal tract.

It is a specific object to provide such delivery system in which theactive pharmacological agent is encapsulated within a protectivematerial which is itself pharmacologically harmless, which does notalter the physiological and biological properties of the active agent,which protects the active agent from the deleterious conditions withinthe gastrointestinal tract and which disappears or releases the activeagent in the bloodstream or other target. It is a further specificobject of this invention to provide such combination of active agent andprotective material which is sufficiently lipophilic and of smallparticle size to pass rapidly through the gastrointestinal mucosa andwhich is simple to manufacture in bulk.

Additional objects of this invention are to provide methods of producingsuch delivery systems and of administering same to animals. It is aspecific object to provide effective means for the oral delivery ofinsulin to diabetic mammals.

It has been found that these objects and other advantages, which will beapparent from this specification, are achieved by the inventiondescribed below.

Broadly, one aspect of this invention is a delivery system for an activepharmacological agent comprising said agent enclosed or encapsulatedwithin proteinoid microspheres.

A second broad aspect of this invention is a method of encapsulating anactive pharmacological agent comprising mixing said active agent with apharmaceutically acceptable liquid and contacting said mixture with aproteinoid that interacts with said mixture to form hollow microspheres.

A third broad aspect of this invention is a method for targeting therelease of a pharmacologically active agent in an animal comprisingadministering to said animal an effective amount of said active agentencapsulated within proteinoid microspheres, said micropheres beingstable to the conditions encountered during migration from the point ofintroduction into said animal to a targeted release zone and beingunstable at said zone.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Proteinoids, which form the protective capsules of this invention, havebeen described as artificial polypeptides, as they are man-madecondensation polymers produced by random or directed assembly of naturalor synthetic aminoacids and/or small peptide chains. Following thediscovery, in the late 1950's, that linear condensation polymers ofmixed natural aminoacids could interact with water to form hollowmicrospheres, proteinoids have been the subject of extensiveinvestigations of the origin of life. An excellent review of theseinvestigations, along with extensive bibliographies, is found in Fox, S.W. and Dose, K., Molecular Evolution and the Origin of Life, MarcelDekker, Inc., New York (1977), the disclosure of which is incorporatedherein by reference.

As a result of these and other studies, a great deal of knowledge hasbeen accumulated concerning the preparation and properties ofproteinoids and proteinoid microspheres. For example, it is known thatproteinoids derived from the natural alpha-aminoacids (those found inanimal or vegetable protein), as well as those incorporating othernaturally occurring materials (such as) polynucleotides, phosphoricacid, iron and calcium), are non-toxic. It also has been found thatinclusion in the polymer of a stoichiometric excess of acidic di orpolycarboxylic aminoacid results in an acidic proteinoid which isinsoluble in an acid environment and soluble in a basic environment,while inclusion of an excess of basic diamino or polyamino monomerresults in a basic proteinoid that is soluble in an acidic medium aandinsoluble at high pH. These solubility characteristics can be veryfinely tuned. Similarly, the size of the microspheres formed bycontacting proteinoids with water or other liquid can be controlledwithin a range of from less that about one half micron to about tenmicrons or more by manipulating a variety of physical or chemicalparameters, such as the pH, osmolarity or salt content of the liquid. Italso has been observed that the proteinoids are far more resistant thanproteins to cleavage by digestive enzymes.

The instant invention arose from the discovery that a pharmacologicallyactive agent can be encapsulated within proteinoid microspheres simplyby dissolving or suspending such agent in a pharmaceutically acceptableliquid, such as water or dimethyl sulfoxide, that interacts with thatproteinoid to form microspheres.

It also has been discovered that such encapsulation does not alter thepharmacological properties of the active agent and that active agentbearing microspheres having diameters of less than about 10 microns aresufficiently small to pass readily through the gastrointestinal mucosaand enter the blood stream. The preferred range for rapid diffusion isfrom about 0.5 to about 5.0 microns, as smaller sizes exhibit somewhatless stability and incorporate relatively little active agent and largersizes diffuse less readily. Particles of from about 5.0 to about 10microns are, however, useful in admixture with those of the preferredrange as their slower diffusion results in prolonged release of theactive agent.

By tailoring both the solubility characteristics of an acidic proteinoidand the particle size of the microspheres by known means, it has beenfound to be possible to produce active agent bearing microspheres whichare stable in the mouth (normal pH of from about 4 to about 6.8), whichrapidly pass through the mucosa of the mouth into the bloodstream andwhich release the active agent in the blood (normal pH of from about7.35 to about 7.45). Such systems are suitable for sublingualadministration of pharmacological agents such as human or bovine growthhormone, interferon or interleukin-II.

Similarly, it is possible to produce readily diffusable microspheresfrom acidic proteinoids which are stable in the highly acidic stomach(normal pH of from about 2 to about 6), but which dissolve in the nearneutral blood. Such systems are suitable for oral administration ofpeptide hormones, such as insulin, or heparin, which otherwise would bequickly destroyed in the stomach. They also are suitable for protectingthe stomach from gastric irritants, such as aspirin. When such aspirincontaining microspheres are orally administered, they pass through thegastrointestinal mucosa and release the aspirin in the bloodstream farmore rapidly than conventional enterically coated aspirin, which firstmust traverse the stomach and then must enter the bloodstream from theintestine after the enteric coating has dissolved.

It also is possible to produce systems from basic proteinoids which arestable in the weakly basic lower digestive tract (normal pH of about 8),but which release active agent in the blood. Such systems are suitablefor the administration of pharmacological agents such as calciumregulators and redox carrier systems for dopamine or gamma-aminobutyricacid.

In addition to these enterically administered delivery systems, it alsois possible to produce a near neutral proteinoid microsphere systemwhich is stable in the bloodstream, but which releases its content ofpharmacological agent in response to the target organ environment, suchas a higher or lower pH or the presence of a specific enzyme. Such nearneutral systems must be introduced intravenously, unless themicrospheres are sufficiently small to be encapsulated within largerproteinoid microspheres that are diffusible through the gastrointestinalmucosa and that are stable until they reach the bloodstream.

Although any pharmacological agent can be encapsulated within proteinoidmicrospheres, it obviously is of particular value for the protection ofsuch agents which otherwise would be destroyed or rendered lesseffective by conditions encountered in the animal body before it reachedits target zone.

Example 1, below, illustrates the preparation of an acidic thermalproteinoid that interacts with an aqueous solution of apharmacologically active agent to encapsulate and protect that agentwithin hollow microspheres. These microspheres exhibit stability in thepresence of the digestive enzymes and acid of the stomach and, beingpredominantly less thin 5.0 microns in diameter, pass readily throughthe gastrointestinal mucosa into the weakly basic blood stream, wherethey dissolve and release the pharmacological agent.

EXAMPLE 1a

A stirred mixture of 52.3 g of aspartic acid (0.4 moles), 42 g ofarginine hydrochloride (0.2 moles), 26 g of isoleucine (0.2 moles) and50 ml of glycerol is heated under nitrogen to 160° C. with the evolutionof gas. The temperature then is maintained at 155° C. for 23 hours,after which the mixture is cooled to room temperature, extracted with200 ml of 10 wt % aqueous sodium bicarbonate and the extract dializedthrough a collodion membrane against distilled water for 26 hours, thewater being changed every six hours. The content of the dialysis tubesthen is evaporated to dryness at 50° C. under vacuum to yield a glassysolid acidic proteinoid material, which is ground to a fine powder.

EXAMPLE 1b

Thirty five mg of this powdered proteinoid is added to a mixture of 50mg of porcine insulin crystals in 2 ml of distilled water and themixture allowed to stand at room temperature until microspheres haveformed. The insulin bearing microspheres are separated by filtration,washed with pH 5.4 aqueous acetic acid and then resuspended in 2 ml ofpH 5.4 aqueous acetic acid. Microscopic examination of this suspensionreveals stable microspheres that are predominantly between 0.1 and 5.0microns in diameter. When a portion of the suspension is neutralized topH 7.4 with concentrated ammonium hydroxide, dissolution of themicrospheres is immediately evident.

EXAMPLE 1c

Each of three adult white rats having normal blood glucose levels isadministered a dose of 0.35 ml of the insulin bearing microspheresuspension of Example 2b by a syringe inserted through the mouth andinto the stomach. Following dosage, each of these animals exhibits asignificant reduction in blood glucose, as measured in blood samplestaken from the tail.

Although hollow microspheres suitable for encapsulating pharmacologicalagents can be formed from proteinoids derived from a single acidic orbasic amino acid and as few as one other amino acid, a greater diversityof component amino acids often produces higher yields of uniform sizemicrospheres within the desirable diameter range of 0.5 to 5.0 microns.Example 2 illustrates the effectiveness, in producing a hypoglycemiceffect in mammals, of the oral administration of insulin encapsulatedwithin a proteinoid derived from 18 different amino acids.

EXAMPLE 2a

A 250 ml filter flask containing 10 g of anhydrous dl-glutamic acid and10 g of anhydrous dl-aspartic acid under nitrogen is heated in an oilbath at approximately 200° C. until the contents are molten. To this isadded 5 g of an anhydrous equimolar mixture of the sixteen neutral andbasic amino acids found in animal protein; i.e., alanine, arginine,asparagine, cysteine, glycine, histadine, leucine, lysine, methionine,phenylalanine, proline, serine, threonine, tyrosine, tryptophan andvaline. The resulting mixture is stirred with a glass rod and maintainedat 200° C. under nitrogen for three hours. After cooling, the amberproduct is extracted with a saturated aqueous solution of sodiumbicarbonate and the resulting solution dialyzed through a collodionmembrane against distilled water at room temperature for 24 hours, thewater being changed every 6 hours. The content of the dialysis tubesthen is acidified to pH 5.4 with concentrated acetic acid andcentrifuged. After discarding the supernatant liquid, the insolublesolids are washed with pH 5.4 aqueous acetic acid and centrifuged again.This wash also is discarded and the solid proteinoid product is driedover silica gel overnight and then ground to a fine powder with mortarand pestle.

EXAMPLE 2b

A mixture of 50 mg of porcine insulin crystals in 2 ml of distilledwater is added to 35 mg of the dry powdered proteinoid of Example 2a andthe mixture allowed to stand at room temperature until microspheres haveformed. The mixture is then centrifuged for 15 minutes. After discardingthe supernatant liquid, the remaining microspheres are washed once withpH 5.4 aqueous acetic acid at room temperature and centrifuged for anadditional 15 minutes. The supernatant liquid again is discarded and theinsulin bearing proteinoid microspheres are resuspended in 2 ml of pH5.4 aqueous acetic acid. Microscopic examination of the suspension showsthat the microspheres are predominantly between 0.5 and 5.0 microns indiameter.

EXAMPLE 2c

Twelve male white rats, each weighing approximately 500 g and having anormal blood glucose level, are randomly assigned to four groups ofthree individuals for demonstrating the physiological efficacy of theoral administration of an aqueous suspension of insulin bearingproteinoid microspheres produced in accordance with the procedure ofExample 2b, above. Between 0.35 and 0.5 ml of this aqueous suspension ofmicrospheres is administered by gavage into the stomach of each rat inGroup One. The Group Two rats have between 1.5 and 1.7 ml of thesuspension similarly administered. The rats of Group Three receive 1.0ml of distilled water similarly administered. The rats of Group Foursimilarly receive 25.0 mg of porcine insulin in 1.0 ml of distilledwater. Both before and during the experiment, all animals are permittedfree access to water and their normal feed. Blood glucose levels aremeasured on samples drawn from the tail at specific intervals aftertreatment and group averages are recorded in Table 1 as milligrams ofglucose per deciliter of blood (mg/dl).

                  TABLE 1                                                         ______________________________________                                        Ave. Blood Glucose (mg/dl)                                                    ______________________________________                                        Group Predose 30 min. 1 hr. 1.5 hr.                                                                             2 hr. 2.5 hr                                                                              3 hr.                           ______________________________________                                        One   135      85      88    66    44    27   --                              Two   --      --      --    --    --     49   --                              Three 120     --      --    --    --    119   --                              Four  113     120     120   124   120   --    116                             ______________________________________                                        Group 4 hr.   6 hr.   12 hr.                                                                              24 hr.                                                                              36 hr.                                                                              48 hr.                                ______________________________________                                        One    38     --      135   119   122   --                                    Two    58     --       80   125   125   122                                   Three 122     --      --    119   124   --                                    Four  111     123     --    --    --    --                                    ______________________________________                                    

It is clear from the data in Table 1 that insulin is delivered in aphysiologically meaningful and active fashion via the oral route withacidic proteinoid microspheres. In all animals receiving insulincontaining microspheres, the blood glucose level returns to thepre-administration level without any observed adverse effect. It shouldbe noted that the administration of larger doses of insulin containingmicrospheres to the Group Two animals appears to increase the durationof action rather than the magnitude of the effect. It also should benoted that the oral administration of far larger dosages per unit ofbody weight of unprotected porcine or bovine insulin to laboratoryanimals and humans produces no detectable reduction of blood glucoselevels.

Similarly effective insulin bearing proteinoid microspheres can beproduced by contacting a dry powered acidic proteinoid, such as those ofExamples 1a or 2a, with insulin suspended or dissolved in a wide varietyof pharmaceutically acceptable liquids, including aqueous solutions ofethanol, isopropanol, terpenol, dimethyl sulfoxide, starch, Tweens 80and cyclodextran.

Example 3 illustrates an especially preferred method of producinginsulin bearing proteinoid microspheres which reliably produces highyields of microspheres that fall within the desirable diameter range of0.5 to 5.0 microns and that are readily soluble at the pH of thetargeted blood.

EXAMPLE 3a

A flask containing 2 parts by weight or anhydrous 1-glutamic acid undera stream of nitrogen is heated in an oil bath at approximately 175° C.until the contents are molten. To this is added 2 parts by weight ofanhydrous 1-aspartic acid and one part by weight of an anhydrousequimolar mixture of the sixteen neutral and basic amino acids found inanimal protein. The resulting mixture is stirred with a glass rod andmaintained at 175° C. under nitrogen for three hours. After cooling, thedark amber product is extracted with saturated aqueous sodiumbicarbonate and the extract dialyzed through a collodion membraneagainst distilled water at room temperature for 24 hours, the waterbeing changed every four hours. The entire content of the dialysis tubesthen is dried under vacuum at 65° and the residual solids are ground toa fine powder with mortar and pestle.

EXAMPLE 3b

An aqueous solution of proteinoid is produced by mixing 35 mg of thepowder of Example 3a per ml of water, adjusting the pH to 7.4 withconcentrated aqueous sodium bicarbonate and removing any insolublematerials by filtration. One part by volume of this solids free solutionof proteinoid then is rapidly injected into an equal volume of a freshlyprepared 25 mg/ml solution of porcine insulin in pH 2.25 aqueous aceticacid. The mixture, which has a pH of approximately 3.5, is stirred in anice bath for 15 minutes and filtered to separate the insulin bearingmicrospheres from the filtrate which is discarded. After washing twicewith pH 3.5 aqueous acetic acid, the microspheres are resuspended in 10parts by volume of pH 3.5 aqueous acetic acid. Microscopic examinationof a portion of this suspension shows a high yield of microspheres whichare predominantly between 0.5 and 5.0 microns in diameter and whichdissolve rapidly when the suspension is neutralized to pH 7.4 by theaddition of concentrated aqueous sodium bicarbonate.

In the following Example 4, dosages of the insulin bearing microspheresuspension of Example 3b are referred to as "insulin filledmicrospheres". Microspheres which contain no encapsulated insulin areproduced by repeating the procedure of Example 3b, except that theinsulin is omitted during the formation of microspheres and themicrospheres are suspended in a 2.5 mg/ml solution of porcine insulin indistilled water, rather than in dilute acetic acid. Doses of theresulting suspension, which contain no insulin within the microspheres,are referred to as "microspheres with external insulin". Dosages of the2.5 mg/ml solution of porcine insulin alone are referred to as "rawinsulin".

EXAMPLE 4

Twelve male white rats, each weighing approximately 500 g and havinng anormal blood glucose level, are arbitrarily assigned to two groups ofthree animals and a third group of six animals. The three animals ofgroup A are administered the insulin filled microspheres by gavage andthe three animals of group B are similarly administered the microsphereswith external insulin. The six animals of group C similarly receive theraw insulin. All dosages are 1 ml/500 g of body weight and all animalsare tested for blood glucose immediately before dosage and at intervalsthereafter. The average blood glucose level for the animals in eachgroup is shown in Table 2.

                  TABLE 2                                                         ______________________________________                                        Blood Glucose (mg/dl) in Rats                                                 Group Treatment                                                                     A            B                                                          Time  (Insulin Filled                                                                            (Microspheres with                                                                          C                                            (hrs) Microspheres)                                                                              External Insulin)                                                                           (Raw Insulin)                                ______________________________________                                        0     109.7        92            92.7                                         .5    54.7         89.3          95.5                                         1     59           84.3          98.2                                         2     50           80.7          95.8                                         3     57.7         86.7          86.2                                         4     64.7         84            91                                           6     76           83.7          89.8                                         8     65.7         88.7          91                                           12    81.7         92.3          92                                           24    94.3         95.7          92                                           ______________________________________                                    

These experiments show no significant effect on blood glucose levels ofeither the raw insulin or the microspheres with external insulin. Incontrast, the insulin filled microspheres produce a peak reduction ofapproximately 50% and an effect of long duration. This demonstrates thatthe acidic proteinoid microspheres have no effect on blood glucoselevels and that they protect only the encapsulated insulin from thehostile environment of the stomach, thereby enabling that encapsulatedinsulin to enter the blood stream in physiologically active form.

EXAMPLE 5a

Diabetes mellitis is induced in rats weighing approximately 300 g bygiving each a 75 mg/kg body weight intravenous injection ofstreptozotocin. Ten rats which are observed to show consistently highblood glucose levels, polyuria and polydipsia and must be maintained onsubcutaneous injections of porcine insulin are selected for thisexperiment.

EXAMPLE 5b

Three of the diabetic rats are administered by gavage approximately 1 mlof the aqueous suspension of porcine insulin bearing acidic proteinoidmicrospheres of Example 3b. A fourth diabetic rat has 3 ml of thesuspension in 50 ml of tap water placed in its water bottle and this ratself administers his dose. All of the rats are food deprived for 12hours prior to dosing. In all subjects, the oral administration of themicroencapsulated insulin produces a significant and prolonged reductionin blood glucose levels.

EXAMPLE 5c

The remaining six diabetic rats are arbitrarily assigned to three groupsof two animals. The animals of the first group are administered bygavage 1 ml of the aqueous suspension of porcine insulin bearing acidicproteinoid microspheres of Example 3b. The animals of the second andthird groups receive subcutaneous injections of 0.25 mg (6.5 I.U.) and0.125 mg (3.25 I.U.), respectively, of porcine insulin. Blood glucosemeasurements are made on all animals immediately before dosing and atintervals thereafter. The animal groups are crossed over twice at oneweek intervals, so that all of the animals receive each of the insulintreatments. The average percent decrease from baseline blood glucoselevels for each treatment is shown in Table 3.

                  TABLE 3                                                         ______________________________________                                        % Decrease from Baseline Blood Glucose in Rats                                        Time (hrs.) after Dosage                                              Insulin Treatment                                                                       0.5   1     1.5 2   3   4   6   8   12  24  48                      ______________________________________                                        Microspheres                                                                            29    38    45  44  48  50  51  40  37  24  -27                     Oral                                                                          0.25 mg SC                                                                              28    55    72  77  84  79  34  4   3   -7  -5                      0.125 mg SC                                                                             5     17    27  45  47  29  8   -8  -8  -8  -8                      ______________________________________                                    

These results demonstrate that the peak effect of the orallyadministered dose of insulin filled microspheres on diabetic rats iscomparable to a subcutaneous injection of 0.125 mg of insulin and thatthe duration of the effect is significantly longer than that produced byeither a 0.125 mg or a 0.25 mg subcutaneous injection.

EXAMPLE 6

One ml of the aqueous suspension of porcine insulin bearing acidicproteinoid microspheres of Example 3b is administered by gavage into thestomach of each of three adult guinea pigs weighing approximately 800 g.Blood samples are taken immediately before and at intervals afteradministration of the dosage.

The blood samples from guinea pig #1 are tested for blood glucose, whichdrops from a predosage level of 160 mg/dl to 42 mg/dl in one half hourand to 25 mg/dl in 1.5 hours, where it remains for an additional 1.5hours, at which time symptoms of insulin shock are observed and theanimal is revived by orally administered glucose.

The blood samples from guinea pigs #2 and #3 are tested for porcineinsulin with radioimmuno assay kits, which are marketed by CambridgeMedical Diagnostics. This method, which distinguishes between porcineand guinea pig insulin, shows that the predosage level of porcineinsulin in the blood of both guinea pigs #2 and #3 is zero. In guineapig #2 the concentration peaks at 250 micrograms/ml one and one halfhours after oral administration of the microspheres and in guinea pig #3a peak of 240 micrograms/ml is reached in four hours.

These experiments demonstrate that the orally administered porcineinsulin has a powerful hypoglycemic effect in a guinea pig, that itactually enters the blood stream and that its administration does notmerely stimulate guinea pig insulin production by the animal.

EXAMPLE 7a

The procedure of Example 3b is repeated, except that the insulin filledmicrospheres are suspended in aqueous acetic acid having a pH of 2.25,rather than 3.5. A sealed vial of this suspension is stored at roomtemperature for 23 days.

EXAMPLE 7b

The activity of the thus aged encapsulated insulin is tested byadministering the suspension by gavage to the stomachs of adult ratswhich have been deprived of rood for eight hours and then measuringblood glucose levels at intervals after dosage. The results are shown inTable 4.

                  TABLE 4                                                         ______________________________________                                        Blood Glucose (mg/dl)                                                                      Time after Dosage (hrs.)                                         Rat #                                                                              weight (g)                                                                             dose (ml)                                                                              0    0.5  1.0  1.5  2.0  3.0                           ______________________________________                                        40   398      0.40     64   49   44   34   33   35                            41   415      0.42     79   89   35   82   80   80                            42   479      0.48     79   50   34   37   37   49                            ______________________________________                                    

Normally insulin in solution can be expected to degrade within a fewdays, even when refrigerated. The magnitude of the blood sugar reductionin all of thes rats and the prolonged effect shown by rats #40 and 42indicate that insulin solution stability is improved by encapsulationwithin acidic proteinoid microspheres.

EXAMPLE 7c

The aged suspension of encapsulated insulin is added to human serum and,employing a standard laboratory haemocytometer counting technique, thenumber of microspheres are counted immediately after mixing and atintervals thereafter. Table 5 shows the number of microspheresobservable as a function of time.

                  TABLE 5                                                         ______________________________________                                        Dissolution of Microspheres in Human Serum                                    ______________________________________                                        Time (minutes)   0     3         30  60                                       Microspheres (× 1000)                                                                    78    50        19  9                                        ______________________________________                                    

This data demonstrates that insulin bearing acidic proteinoidmicrospheres which have remained intact after 23 days exposure at roomtemperature to pH 2.25 aqueous acetic acid still dissolve rapidly innear neutral human serum.

EXAMPLE 8a

An aqueous solution of heparin containing 250 mg/ml of heparin isadjusted to pH 4.5 by the addition of concentrated acetic acid. To thisis added 35 mg/ml of the dry powdered acidic proteinoid of Example 3aand the mixture is allowed to stand at room temperature untilmicrospheres have formed. One part by volume of the mixture then iscentrifuged and, after discarding the supernatant liquid, the heparinbearing microspheres are washed with pH 4.5 aqueous acetic acid,filtered and resuspended in pH 4.5 aqueous acetic acid, the suspensionbeing made up to one part by volume. Microscopic examination revealsthat the microspheres are predominantly within the range of from about0.1 to about 5 microns in diameter, with the majority being between 1and 2 microns.

EXAMPLE 8b

Seven male white rats, each weighing approximately 600 g, are deprivedof food for 12 hours before the start of the experiment. Rat #1 receivesno treatment. Rat #2 receives an intravenous injection of 250 mg ofheparin in one ml of distilled water. Each of rats #3-7 has 1 ml of theaqueous suspension of heparin bearing microspheres of Example 8aintroduced directly into the stomach by gavage. The effect of heparin isdetermined using the Activated Partial Thromboplastin Time (APTT) test.This test measures the time necessary for a sample of serum taken fromthe tail vein to form a fibrin clot. The results for each rat at varioustimes after dosing are shown in Table 6.

                  TABLE 6                                                         ______________________________________                                        Time to Coagulation in the APTT Test (seconds)                                     Heparin                                                                  Rat #                                                                              Treatment Predose 1 hr. 2 hrs.                                                                             3 hrs.                                                                              4 hrs.                                                                             24 hrs.                          ______________________________________                                        1    none      28      26    27   --    --   --                               2    IV        25      54    >300 --    --   --                               3    microspheres                                                                            24      35    >300 --    --   --                               4    "         26      35    59   118   >300 21                               5    "         29      41    63   106   248  26                               6    "         25      37    66   121   >300 23                               7    "         24      31    62   111   >300 37                               ______________________________________                                    

In all animals receiving heparin bearing microspheres, coagulation timeincreases to a level comparable to that seen following an intravenousinjection of heparin. It is clear from this data that heparin isdelivered to the bloodstream in a physiologically meaningful and activefashion when encapsulated in acidic proteinoid microspheres andadministered orally. It should be noted that the oral administration offar larger doses per unit of body weight of unprotected heparin tolaboratory animals and humans produces no detectable increase incoagulation time.

EXAMPLE 9a

An aqueous solution of physostigmine containing 50 mg/ml ofphysostigmine is adjusted to pH 5 by the addition of concentrated aceticacid. To one volume of this solution is added 100 mg per ml of the drypowdered acidic proteinoid of Example 3a and the mixture is allowed tostand at room temperature until microspheres have formed. It then isfiltered, washed three times with pH 5 aqueous acetic acid and theseparated microspheres are resuspended in one volume of pH 5 aceticacid. Microscopic examination reveals that the suspended microspheresare predominantly 0.5-5.0 microns in diameter.

EXAMPLE 9b

Each of two normal rats weighing about 360 g is administered by gavage 3ml of the suspension of physostigmine bearing microspheres. Within 30minutes of dosage, both animals have died and each exhibits enlargedliver and peritoneal hemorrhage. These lethal oral dosages ofmicroencapsulated physostigmine are calculated to be less than onepercent of the LD₅₀ oral dosage of unprotected physostigmine in rats.

In addition to the specific pharmacological agents that are shown by theabove examples to be released in the blood stream in physiologicallyactive form when orally administered within protective acidic proteinoidmicrospheres, such microsphere delivery system is similarly effectivewith a wide variety of other agents which are labile in the stomachinvironment, including nitroglycerin, Salk polio vaccine, rubellavaccine and hepatitis B vaccine. There are, however many otherpharmacological agents which could be deleteriously affected by even themildly acidic conditions encountered during encapsulation within acidicproteinoid microspheres.

The following experiment demonstrates the ability of a basic proteinoidto form microspheres which encapsulate and protect one such extremelyacid sensitive pharmacological agent, a dopamine derivative, from thehostile environment of the gastrointestinal tract, as well as to deliverthat agent to the circulatory system, from which it penetrates the brainblood barrier and releases dopamine in the brain. The dopaminederivative employed in this experiment is PR-21, which is a proprietarycomposition of acylated dopamine bonded to a reduceddihydropyridine/pyridinium salt type redox carrier which was developedby Pharmatek, Inc. and is described in U.S. Pat. No. 4,479,932. Theunprotected PR-21 composition is unstable anywhere in thegastrointestinal tract and is particularly sensitive to acid conditions.When injected intravenously into rats, significant amounts of thedeacylated quaternary precursor of dopamine can be measured in thehomogenized rat brain by the method of Bodir and Farog, Journal ofMedicinal Chemistry, 26, 528 (1983).

EXAMPLE 10a

A nitrogen swept mixture of two parts by weight of arginine, two partsby weight of lysine and one part by weight of an equimolar mixture ofthe sixteen neutral and acidic aminoacids found in animal protein isstirred and heated at 180° C. for 3 hours. The cooled reaction mixtureis extracted with pH 2.25 aqueous acetic acid and the extract isdialyzed through a collodion membrane against a large volume ofdistilled water at room temperature for 48 hours, the water beingchanged every six hours. The content of the dialysis tubes then isheated under vacuum at 65° C. to yield a dry powdered basic proteinoid.When suspended in a moderate to strongly alkaline liquid environment,this powdered proteinoid spontaneously forms hollow microspheres whichare stable in that environment, but which dissolve at the near neutralpH of blood.

EXAMPLE 10b

One part by volume of an ethanol solution of PR-21 (360 mg/ml) isdiluted with an equal volume of distilled water and the pH of thesolution is adjusted to 8 by the addition of saturated aqueous monobasicpotassium phosphate buffer. A portion of this buffered solution, whichcontains 180 mg/ml of PR-21, is set aside and dosages of it are referredto below as "unprotected PR-21".

The remainder of the buffered solution is mixed with 25 mg/ml of the drypowdered basic proteinoid of Example 10a and chilled in an ice bathuntil microspheres have formed. Dosages of the resulting suspension, inwhich the microspheres are predominantly 0.1 to 5 microns in diameter,are referred to below as "microencapsulated PR-21".

EXAMPLE 10c

Two rats weighing about 500 g (rats DA-1 and DA-2) are anesthetized, thejejunum is externalized and the sphincter is tied off to preventbackwash into the stomach. Two ml of microencapsulated PR-21 are theninjected into the jejunum of each rat. Two similar control rats (ratsDA-5 and DA-6) are similarly prepared, but are injected in the jejunumwith 2 ml of unprotected PR-21. Finally, two similar control rats (ratsDA-3 and DA-4) are intravenously injected with 2 ml of unprotectedPR-21. Table 7 shows the amount of deacylated quaternary precursor ofdopamine that is detectable in the homogenized brains of the sixsubjects.

                  TABLE 7                                                         ______________________________________                                        Dopamine Quaternary Precursor in Rat Brains                                   Rat #   Treatment       Precursor (micrograms/g)                              ______________________________________                                        DA-1    Intestinal Injection,                                                                         1                                                             Microencapsulated PR-21                                               DA-2    Intestinal Injection,                                                                         10                                                            Microencapsulated PR-21                                               DA-3    IV Injection,   3                                                             Unprotected PR-21                                                     DA-4    IV Injection,   4.5                                                           Unprotected PR-21                                                     DA-5    Intestinal Injection,                                                                         0                                                             Unprotected PR-21                                                     DA-6    Intestinal Injection,                                                                         0                                                             Unprotected PR-21                                                     ______________________________________                                    

These results demonstrate the capacity of basic proteinoid microspheresto encapsulate and protect a dopamine derivative from the digestiveenzymes and basic environment of the intestine, as well as the fact thatsuch microspheres are transported across the gastrointestinal mucosainto the rear neutral blood stream where the encapsulated dopaminederivative is released. For successful oral delivery of suchencapsulated pharmacological agent, the acid sensitive basic proteinoidmicrospheres must be protected while traversing the mouth and stomach.Advantageously, this is accomplished by a conventional enteric coatingwhich does not dissolve until it reaches the intestine.

EXAMPLE 11

A stirred mixture of 2 mole parts of anhydrous glutamic acid, 2 moleparts of lysine and 1 mole part of an equimolar mixture of neutralaminoacids (alanine, glycine, leucine, phenylalanine, proline, tyrosineand valine) is heated under nitrogen at 170° C. for four hours. Thecooled reaction product is extracted with pH 2.25 aqueous acetic acidand the extract dialyzed through a collodion membrane against distilledwater for 24 hours, the water being changed every 4 hours. The contentof the dialysis tubes is evaporated to dryness at 65° C. under vacuumand the residual solids are ground to a fine powder. When added to a pH7.4 aqueous solution or suspension of a pharmacological agent, thisneutral powdered proteinoid spontaneously forms a profusion of hollowmicrospheres which encapsulate that solution or suspension.

These microspheres are stable in human serum, but dissolve rapidly in pH2.5 aqueous acid to release their contents. Being destabitized byexposure to reduced pH, such as that encountered when engulfed withinmacrophages, these neutral proteinoid microspheres are suited for theintravenous adminstration of a pharmacological agent, such asazidothymidine, which, in unprotected form, is quickly absorbed by manyuntargeted body tissues and cells, as well as the targeted macrophages.

It will be apparent to those of ordinary skill in the art that numerouschanges and modifications can be made in the illustrative embodiments ofthe invention described above without departing from the spirit or scopeof the invention as set forth in the following claims.

We claim:
 1. Composition comprising a pharmacologically active agentencapsulated within protenoid microspheres having diameterspredominantly less than about 10 microns and formed from linear thermalcondensation polymers of mixed amino acids.
 2. Composition of claim 1wherein said microspheres are stable in at least a segment of thegastrointestinal tract, are unstable in the blood stream and arepredominatly less than about 10 microns in diameter so as to readilypenetrate the gastrointestinal mucosa and release said active agent inthe blood stream in physiologically active form.
 3. Composition of claim2 wherein said polymer is acidic and said microspheres are stable toacids and enzymes in the mouth.
 4. Composition of claim 2 wherein saidpolymer is basic and said microspheres are stable in the weakly basiclower digestive tract.
 5. Composition of claim 4 wherein saidpharmacological agent is a dopamine redox carrier system.
 6. Compositionof claim 2 wherein said polymer is acidic and said microspheres arestable to acids and enzymes in the stomach.
 7. Composition of claim 6wherein said pharmacological agent is insulin.
 8. Composition of claim 6wherein said pharmacological agent is heparin.
 9. Composition of claim 6wherein said pharmacological agent is physostigmine.
 10. Composition ofclaim 1 wherein said polymer is neutral and said microspheres are stablein the blood stream and are unstable at reduced pH.
 11. Composition ofclaim 1 wherein said microspheres are predominantly from about 0.5 toabout 5.0 microns in diameter.
 12. Method for microencapsulating apharmacologically active agent within microspheres for targeted releasewithin a selected pH range comprising forming a mixture of said agentwith a pharmaceutically acceptable liquid, said mixture having a pHoutside said selected range, and contacting said mixture withproteinoids formed of linear thermal condensation polymers of mixedamino acids which are soluble within said selected pH range andinsoluble in said mixture to form microspheres having diameterspredominately less than about 10 microns containing the active agent.13. Method of claim 12 wherein said pharmaceutically acceptable liquidis water.
 14. Method of claim 12 including preliminary purification ofsaid polymers by mixing the polymers with water having a pH within saidselected range and separating the resulting aqueous solution of saidpolymers from any insoluble material.
 15. Method for producing an orallyadministerable composition for delivering insulin to the blood stream inphysiologically active form comprising mixing insulin with water andcontacting said mixture with a thermal condensation polymer derived fromabout two parts glutamic acid, about two parts aspartic acid and aboutone part of neutral or basic alpha-aminoacid.
 16. Method for targetingrelease of a pharmacologically active agent in an animal comprisingadministering to said animal an effective amount of said active agentencapsulated within proteinoid microspheres formed from linear thermalcondensation polymers of mixed amino acids having diameterspredominately less than about 10 microns, said microspheres being stableto the conditions encountered during migration from the point ofintroduction into said animal to a targeted release zone and beingunstable at said zone.
 17. Method of claim 16 wherein said microspheresare predominantly from about 0.5 to about 5.0 microns in diameter. 18.Method of claim 16 wherein said microspheres are entericallyadministered for targeted release in the blood stream and saidmicrospheres are stable in that segment of the gastrointestinal tractinto which they are introduced and are unstable in the blood stream. 19.Method of claim 18 wherein said encapsulated active agent issublingually introduced into the bloodstream and said microspheres aresufficiently acidic to be stable at a pH of from about 4 to about 6.8and unstable at a pH of from about 7.35 to about 7.45.
 20. Method ofclaim 18 wherein said encapsulated active agent is introduced into thebloodstream from the weakly basic lower gastrointestinal tract and saidmicrospheres are sufficiently basic to be stable at a pH of about 8 andunstable at a pH of from about 7.35 to about 7.45.
 21. Method of claim18 wherein said encapsulated active agent is gastrically introduced intothe bloodstream and said microspheres are sufficiently acidic to bestable at a pH of from about 2 to about 6 and unstable at a pH of fromabout 7.35 to about 7.45.
 22. Method of claim 16 wherein saidmicrospheres are intravenously administered and are stable at a pH offrom about 7.35 to about 7.45 and unstable at reduced pH.
 23. Method oftreating a diabetic conditions in an animal comprising orallyadministering to said animal an effective amount of insulin which isencapsulated within acidic protenoid microspheres formed from linearthermal condensation polymers of mixed amino acids, said microspheresbeing predominantly from about 0.5 to about 5.0 microns in diameter,stable at a pH of up to about 6 and unstable at a pH of from about 7.35to about 7.45. .Iadd.
 24. Composition comprising a pharmacologicallyactive agent encapsulated within proteinoid microspheres havingdiameters predominantly less than about 10 microns and formed fromthermal condensation polymers of mixed amino acids..Iaddend..Iadd.25.Method for microencapsulating a pharmacologically active agent withinmicrospheres for targeted release within a selected pH range comprisingforming a mixture of said agent with a pharmaceutically acceptableliquid, said mixture having a pH outside said selected range, andcontacting said mixture with proteinoids formed of thermal condensationpolymers of mixed amino acids which are soluble within said selected pHrange and insoluble in said mixture to form microspheres havingdiameters predominantly less than about 10 microns containing the activeagent..Iaddend..Iadd.26. Method for targeting release of apharmacologically active agent in an animal comprising administering tosaid animal an effective amount of said active agent encapsulated withinproteinoid microspheres formed from thermal condensation polymers ofmixed amno acids having diameters predominately less than about 10microns, said microspheres being stable to the conditions encounteredduring migration from the point of introduction into said animal to atargeted release zone and being unstable at said zone..Iaddend..Iadd.27.Method of treating diabetic conditions in an animal comprising orallyadministering to said animal an effective amount of insulin which isencapsulated within acidic proteinoid microspheres formed from thermalcondensation polymers of mixed amino acids, said microspheres beingpredominantly from about 0.5 to about 5.0 microns in diameter, stable ata pH of up to about 6 and unstable at a pH of from about 7.35 to about7.45..Iaddend..Iadd.28. Composition comprising a pharmacologicallyactive agent encapsulated within proteinoid microspheres havingdiameters predominantly less than about 10 microns and formed fromcondensation polymers of mixed amino acids..Iaddend..Iadd.29. Method formicroencapsulating a pharmacologically active agent within microspheresfor targeted release within a selected pH range comprising forming amixture of said agent with a pharmaceutically acceptable liquid, saidmixture having a pH outside said selected range, and contacting saidmixture with proteinoids formed of condensation polymers of mixed aminoacids which are soluble within said selected pH range and insoluble insaid mixture to form microspheres having diameters predominantly lessthan about 10 microns containing the active agent..Iaddend..Iadd.30.Method for targeting release of a pharmacologically active agent in ananimal comprising administering to said animal an effective amount ofsaid active agent encapsulated within proteinoid microspheres formedfrom condensation polymers of mixed amino acids having diameterspredominately less than about 10 microns, said microspheres being stableto the conditions encountered during migration from the point ofintroduction into said animal to a targeted release zone and beingunstable at said zone..Iaddend..Iadd.31. Method of treating diabeticconditions in an animal comprising orally administering to said animalan effective amount of insulin which is encapsulated within acidicproteinoid microspheres formed from condensation polymers of mixed aminoacids, said microspheres being predominantly from about 0.5 to about 5.0microns in diameter, stable at a pH of up to about 6 and unstable at apH of from about 7.35 to about 7.45..Iaddend..Iadd.32. Compositioncomprising a pharmacologically active agent encapsulated withinproteinoid microspheres having diameters predominantly less than about10 microns and formed from polymers of mixed aminoacids..Iaddend..Iadd.33. Method for microencapsulating apharmacologically active agent within microspheres for targeted releasewithin a selected pH range comprising forming a mixture of said agentwith a pharmaceutically acceptable liquid, said mixture having a pHoutside said selected range, and contacting said mixture withproteinoids formed of polymers of mixed amino acids which are solublewithin said selected pH range and insoluble in said mixture to formmicrospheres having diameters predominantly less than about 10 micronscontaining the active agent..Iaddend..Iadd.34. Method for targetingrelease of a pharmacologically active agent in an animal comprisingadministering to said animal an effective amount of said active agentencapsulated within proteinoid microspheres formed from polymers ofmixed amino acids having diameters predominately less than about 10microns, said microspheres being stable to the conditions encounteredduring migration from the point of introduction into said animal to atargeted release zone and being unstable at said zone..Iaddend..Iadd.35.Method of treating diabetic conditions in an animal comprising orallyadministering to said animal an effective amount of insulin which isencapsulated within acidic proteinoid microspheres formed from polymersof mixed amino acids, said microspheres being predominantly from about0.5 to about 5.0 microns in diameter, stable at a pH of up to about 6and unstable at a pH of from about 7.35 to about 7.45..Iaddend..Iadd.36.Composition comprising a pharmacologically active agent encapsulatedwithin proteinoid microspheres having diameters predominantly less thanabout 10 microns and formed from mixed amino acids..Iaddend..Iadd.37.Method for microencapsulating a pharmacologically active agent withinmicrospheres for targeted release within a selected pH range comprisingforming a mixture of said agent with a pharmaceutically acceptableliquid, said mixture having a pH outside said selected range, andcontacting said mixture with proteinoids formed of mixed amino acidswhich are soluble within said selected pH range and insoluble in saidmixture to form microspheres having diameters predominantly less thanabout 10 microns containing the active agent..Iaddend..Iadd.38. Methodfor targeting release of a pharmaceutically active agent in an animalcomprising administering to said animal an effective amount of saidactive agent encapsulated within proteinoid microspheres formed frommixed amino acids having diameters predominately less than about 10microns, said microspheres being stable to the conditions encounteredduring migration from the point of introduction into said animal to atargeted release zone and being unstable at said zone..Iaddend..Iadd.39.Method of treating diabetic conditions in an animal comprising orallyadministering to said animal an effective amount of insulin which isencapsulated within acidic proteinoid microspheres formed from mixedamino acids, said microspheres being predominantly from about 0.5 tabout 5.0 microns in diameter, stable at a pH of up to about 6 andunstable at a pH of from about 7.35 to about 7.45..Iaddend..Iadd.40.Composition comprising a pharmacologically active agent encapsulatedwithin proteinoid microspheres having diameters predominantly less thanabout 10 microns and formed from directed polymers of mixed aminoacids..Iaddend..Iadd.41. Method for microencapsulating apharmacologically active agent within microspheres for targeted releasewithin a selected pH range comprising forming a mixture of said agentwith a pharmaceutically acceptable liquid, said mixture having a pHoutside said selected range, and contacting said mixture withproteinoids formed of directed polymers of mixed amino acids which aresoluble within said selected pH range and insoluble in said mixture toform microspheres having diameters predominantly less than about 10microns containing the active agent..Iaddend..Iadd.42. Method fortargeting release of a pharmacologically active agent in an animalcomprising administering to said animal an effective amount of saidactive agent encapsulated within proteinoid microspheres formed fromdirected polymers of mixed amino acids having diameters predominatelyless than about 10 microns, said microspheres being stable to theconditions encountered during migration from the point of introductioninto said animal to a targeted release zone and being unstable at saidzone..Iaddend..Iadd.43. Method of treating diabetic conditions in ananimal comprising orally administering to said animal an effectiveamount of insulin which is encapsulated within acidic proteinoidmicrospheres formed from directed polymers of mixed amino acids, saidmicrospheres being predominantly from about 0.5 to about 5.0 microns indiameter, stable at a pH of up to about 6 and unstable at a pH of fromabout 7.35 to about 7.45..Iaddend..Iadd.44. Composition comprising apharmacologically active agent encapsulated within proteinoidmicrospheres having diameters predominantly less than about 10 micronsand formed from directed condensation polymers of mixed aminoacids..Iaddend..Iadd.45. Method for microencapsulating apharmacologically active agent within microspheres for targeted releasewithin a selected pH range comprising forming a mixture of said agentwith a pharmaceutically acceptable liquid, said mixture having a pHoutside said selected range, and contacting said mixture withproteinoids formed of directed condensation polymers of mixed aminoacids which are soluble within said selected pH range and insoluble insaid mixture to form microspheres having diameters predominantly lessthan about 10 microns containing the active agent..Iaddend..Iadd.46.Method for targeting release of a pharmacologically active agent in ananimal comprising administering to said animal an effective amount ofsaid active agent encapsulated within proteinoid microspheres formedfrom directed condensation polymers of mixed amino acids havingdiameters predominately less than about 10 microns, said microspheresbeing stable to the conditions encountered during migration from thepoint of introduction into said animal to a targeted release zone andbeing unstable at said zone..Iaddend..Iadd.47. Method of treatingdiabetic conditions in an animal comprising orally administering to saidanimal an effective amount of insulin which is encapsulated withinacidic proteinoid microspheres formed from directed condensationpolymers of mixed amino acids, said microspheres being predominantlyfrom about 0.5 to about 5.0 microns in diameter, stable at a pH of up toabout 6 and unstable at a pH of from about 7.35 to about7.45..Iaddend..Iadd.48. Composition comprising a pharmacologicallyactive agent encapsulated within proteinoid microspheres havingdiameters predominantly less than about 10 microns and formed fromrandom polymers of mixed amino acids..Iaddend..Iadd.49. Method formicroencapsulating a pharmacologically active agent within microspheresfor targeted release within a selected pH range comprising forming amixture of said agent with a pharmaceutically acceptable liquid, saidmixture having a pH outside said selected range, and contacting saidmixture with proteinoids formed of random polymers of mixed amino acidswhich are soluble within said selected pH range and insoluble in saidmixture to form microspheres having diameters predominantly less thanabout 10 microns containing the active agent..Iaddend..Iadd.50. Methodfor targeting release of a pharmacologically active agent in an animalcomprising administering to said animal an effective amount of saidactive agent encapsulated within proteinoid microspheres formed fromrandom polymers of mixed amino acids having diameters predominately lessthan about 10 microns, said microspheres being stable to the conditionsencountered during migration from the point of introduction into saidanimal to a targeted release zone and being unstable at saidzone..Iaddend..Iadd.51. Method of treating diabetic conditions in ananimal comprising orally administering to said animal an effectiveamount of insulin which is encapsulated within acidic proteinoidmicrospheres formed from random polymers of mixed amino acids, saidmicrospheres being predominantly from about 0.5 to about 5.0 microns indiameter, stable at a pH of up to about 6 and unstable at a pH of fromabout 7.35 to about 7.45..Iaddend..Iadd.52. Composition comprising apharmacologically active agent encapsulated within proteinoidmicrospheres having diameters predominantly less than about 10 micronsand formed from random condensation polymers of mixed aminoacids..Iaddend..Iadd.53. Method for microencapsulating apharmacologically active agent within microspheres for targeted releasewithin a selected pH range comprising forming a mixture of said agentwith a pharmaceutically acceptable liquid, said mixture having a pHoutside said selected range, and contacting said mixture withproteinoids formed of random condensation polymers of mixed amino acidswhich are soluble within said selected pH range and insoluble in saidmixture to form microspheres having diameters predominantly less thanabout 10 microns containing the active agent..Iaddend..Iadd.54. Methodfor targeting release of a pharmacologically active agent in an animalcomprising administering to said animal an effective amount of saidactive agent encapsulated within proteinoid microspheres formed fromrandom condensation polymers of mixed amino acids having diameterspredominately less than about 10 microns, said microspheres being stableto the conditions encountered during migration from the point ofintroduction into said animal to a targeted release zone and beingunstable at said zone..Iaddend..Iadd.55. Method of treating diabeticconditions in an animal comprising orally administering to said animalan effective amount of insulin which is encapsulated within acidicproteinoid microspheres formed from random condensation polymers ofmixed amino acids, said microspheres being predominantly from about 0.5to about 5.0 microns in diameter, stable at a pH of up to about 6 andunstable at a pH of from about 7.35 to about 7.45..Iaddend.